ANANDAMIDE/THC INDUCED ARACHIDONIC ACID RELEASE

本申请中提出的实验的目标是确定 导致大麻素诱导的花生四烯酸的生化事件 release.将要进行的研究将集中于信号 大麻素与其受体结合后的转导过程。的 在这一过程中受体介导的问题将由 反义寡脱氧核苷酸的用途 常规配体结合方法。寡核苷酸将被设计为减少， 或完全消除大麻素受体基因的表达 并与随后花生四烯酸释放的变化进行比较。术语 大麻素包括外源性物质，如THC， 和内源性分子如大麻素用于这些的系统 研究将分离完整的细胞，如小鼠腹膜 巨噬细胞、神经母细胞瘤和中国仓鼠卵巢系。的 实验旨在确定磷脂酶D，蛋白质 激酶C、MAP激酶和磷脂酶A2介导大麻素- 刺激花生四烯酸的释放和随后的 类花生酸酶活性的直接测量将在 大麻素浓度范围，并与花生四烯酸比较 在相同条件下释放。还将进行时间进程研究 以帮助阐明所涉及的事件的顺序。的特异性抑制剂 在可能的情况下，将使用各种酶来补充数据 通过直接测量获得。 类花生酸，如前列腺素E2，已被证明是重要的 在人类和动物模型中， 详细了解它们的水平是如何升高的， 重要性最近发现的内源性大麻素，大麻酰胺， 可能作为大脑神经调节器发挥作用， 由于其结构性， 天然产物，即花生四烯酸的乙醇肽衍生物。的 大麻素的发现对“前列腺素假说”的意义 大麻素的作用目前尚不清楚，但是， 花生四烯酸的类花生酸性质只是巧合。更好的 了解THC/anandamide的生化作用可以提高我们的 对记忆、运动协调等过程的理解 能力、时间感和自我知觉。此外，一些见解可能 因此，在大麻滥用的分子基础的问题上，的 最近关于大麻使用日益增加的报告， 青春期前，再次将注意力集中在努力了解 这种药物的作用。

The goal of the experiments proposed in this application is to determine the biochemical events that lead to cannabinoid-induced arachidonic acid release. The studies to be carried out will be focused on the signal transduction processes following cannabinoid binding to its receptor. The question of receptor mediation in this process will be addressed by the use of anti sense oligodeoxynucleotides in addition to the more conventional ligand binding approach. Oligos will be designed to reduce, or eliminate completely, the expression of the cannabinoid receptor genes and compared with subsequent changes in arachidonic acid release. The term cannabinoid is meant to include both exogenous substances, such as THC, and endogenous molecules like anandamide. The systems to be used for these investigations will be isolated, intact cells such as mouse peritoneal macrophages, neuroblastoma and Chinese-hamster ovary lines. The experiments are aimed at defining roles for phospholipase D, protein kinase C, MAP kinase and phospholipase A2 in mediating cannabinoid- stimulated release of arachidonic acid and the subsequent production of eicosanoids. Direct measurement of enzyme activities will be made over a range of cannabinoid concentrations and compared with arachidonic acid release under identical conditions. Time course studies will also be done to help elucidate the sequence of events involved. Specific inhibitors of the various enzymes will be used where possible to supplement the data obtained by direct measurement. Eicosanoids, such as prostaglandin E2, have been shown to be important mediators of cannabinoid action in humans as well as in animal models so that a detailed understanding of how their levels are elevated is of importance. The recent discovery of an endogenous cannabinoid, anandamide, that may function as a brain neuroregulator, lends even greater significance to the aims of this proposal because of its structural nature, namely, the ethanolantide derivative of arachidonic acid. The significance of the anandamide discovery to the "prostaglandin hypothesis" of cannabinoid action is not clear at this time, however, it is unlikely that the eicosanoid nature of anandamide is only coincidental. A better understanding of THC/anandamide biochemical actions could improve our understanding of processes such as memory, motor coordination cognitive ability, time sense and self-perception. In addition, some insight might be gained into the question of the molecular basis for cannabis abuse. The recent reports on the increasing use of marihuana, especially among preadolescents, has again focussed attention on efforts to understand the actions of this drug.